ANTIMICROBIAL EFFECTS OF HONEY ON SOME FUNGAL PATHOGENS

ABSTRACT

The antimicrobial effect of some honey samples obtained from National Root Crop Research Institute (NRCRI), Umudike and Mbaise were tested on some fungal pathogens such as Aspergillus niger, Penicillium chrysogenum and Candida albicans. The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the honeys were also determined. Results obtained reveal that the honey samples showed varying levels of inhibitory activity at various concentrations against the fungi tested with zones of inhibition increasing with increasing honey concentration. Penicillium chrysogenum was the most sensitive of all the fungal isolates studied, value at 21mm at 100% in honey from National Root Crop Research Institute (NRCRI), Umudike and 22mm at 100% in honey from Mbaise. While C. albicans was the least sensitive, value at 18mm at 100% in honey from National Root Crop Research Institute (NRCRI), Umudike and 19mm in honey from Mbaise. The minimum inhibitory concentration (MIC) value from 40 – 60% concentration while the minimum fungicidal concentration (MFC) value from 20 – 40% concentration. The samples of honey used in the study showed broad spectrum and promising antifungal effect. Nigerian honey can serve as sources of antifungal substances for possible development of antifungal drugs for the treatment of fungal infections.  

TABLE OF CONTENTS

Title page         i

Certification         ii

Dedication         iii

Acknowledgements         iv

Table of contents         v

List of tables         ix

Abstract         x

CHAPTER ONE

1.1 Introduction          1

1.2 Aims and Objectives          2

CHAPTER TWO

2.1 Literature Review           3

2.2 Local test for real Honey           5

2.2.1 Classification of  Honey           5

2.2.2 Extraction           6

2.2.3 Preservation           6

2.3 Properties and active ingredients of honey           7

2.4 Mode of action of some of the antibacterial substances in honey                    8

2.4.1 High osmotic pressure                                                                       8

2.4.2 Low water activity           8

2.4.3 Glucose oxidase enzyme           8

2.4.4 Low pH/ acidic environment           9

2.5 Clinical conditions that respond to treatment with honey                9

2.6 Honey as an antibacterial agent           10

2.6.1 Practical consideration for the clinical use of honey         12

2.6.2 Adverse reaction of honey         12

2.7 Nutritional and health benefits of honey         13

2.7.1 Health         13

2.7.2 Gastroenterology                     13

2.7.3 Infants         14

2.8 Fungal pathogens                     15

2.8.1 Aspergillus niger         15

2.8.2 Uses of Aspergillus niger         15

2.8.3 Penicillium chrysogenum         15

2.8.4 Uses of Penicillium chrysogenum         16

2.8.5 Candida albicans         16

CHAPTER THREE

3.0 Material and methods         17

3.1 Material         17

3.2 Methods         17

3.2.1 Collection of samples         17

3.2.2 Microorganisms used           17

3.2.3 Honey preparation         18

3.2.4 Media used         18

3.2.5 Preparation of Inoculum         18

3.3 Determination of antifungal activities           18

3.4 Determination of minimum inhibitory concentration (MIC)           19

3.5 Determination of minimum fungicidal concentration (MFC)         19

CHAPTER FOUR

4.1 Results           21

CHAPTER FIVE

5.1 Discussion            26

5.2 Conclusion                28

REFERENCES         29

LIST OF TABLES

TABLES TITLE PAGE

1:    The zone of inhibition on the effect of undiluted honey on some

        fungal pathogens.         22

2:     Antimicrobial effect of honey sample on the test organisms.         23

3:       Minimum inhibitory concentration of the honey samples

                    against test organisms.         24

4:     Minimum fungicidal concentration of the honey samples

                 against test organisms.         25

CHAPTER ONE

1.1 INTRODUCTION

Honey is a natural sweet substance produced by honeybees from the nectar of blossoms or from the secretions of living parts of plants or excretions of plant- sucking insects on the living parts of plants, which honeybees collect, transform, and combine with specific substances of their own, store, and leave in the honeycomb to ripen and mature (Alimentarius., 2001). It has been reported that honey contains carbohydrates, mainly fructose and glucose in addition to about 25 different oligosaccharides (Bogdanov et al., 2008). Honey possesses powerful antimicrobial properties that can be utilized at low cost and at no risk (Fessenden., 2008). Various studies have reported the antimicrobial effect of honey (Agbaje et al., 2006; Portillo et al., 2001; Vilma et al., 2007). Honey has been shown by (Mekky., 2007) to inhibit the growth of Aspergillus flavus and reduce aflatoxin B1 and B2 levels. The intrinsic properties of honey have been reported to affect the growth and survival of microorganisms by bacteriostatic or bactericidal actions (Iurlina and Fritz., 2005). Although several in vitro studies have demonstrated the antibacterial properties of honey (Lusby et al., 2005; Kwakman, 2008; Hassanain et al., 2010), few have examined  the  action  against  fungi.  The incidence of fungal infections is increasing in both the community and hospital environments with several causative agents including yeasts with Candida spp., among the leading organisms (Abi-Said et al., 1997; Pfaller and Diekema., 2002) and filamentous fungi. Antifungal action of honey has been observed against the yeast Candida albicans and most species of Aspergillus baumannii and Penicillium chrysogenum (Willix et al., 1992) as well as all the common dermatophytes (Brady et al., 1997). The production and type of honey produced by honeybees is dependent on the natural vegetative flowers blooming in different seasons. Thus, the flowers from which bees gather nectar to produce honey may contribute to the difference in the antimicrobial effect. However, large variations in the in-vitro antibacterial activity of various types of honey have been reported and thus hampered its acceptance in modern medicine (Kwakman., 2008). Honey is farmed and used in several parts of Nigeria. Initially, local farmers harvested the honey from the wild but today, apiculture is being practiced in many parts of the country. Whereas there are large volumes of data on the biological activities of honeys from different parts of the world, including North America, Europe, Asia, Australia, and South Africa, there is a dearth of data on the biological activities of Nigerian honeys. Although some researchers (Adesunkanmi and Oyelami., 1994; Omafuvbe and Akanbi, 2009) have reported the antibacterial effect of honey collected from Nigeria, information about the antifungal effect of Nigerian honey are still scarce.

1.2 AIMS AND OBJECTIVES

To determine the inhibitory activities of honey on fungal isolates (Candida albicans, Aspergillus niger and Penicillium chrysogenum,).

To determine the Minimum Inhibitory Concentration (MIC) of honey on Candida albicans, Aspergillus niger and Penicillium chrysogenum.

To determine the Minimum fungicidal Concentration (MFC) of honey on Candida albicans, Aspergillus niger and Penicillium chrysogenum .

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